| Objective:The purpose of this study was to explore the influence of key factors during the cultivation process on the quality of Paeoniae Radix Alba using Q-Marker as quality control standard.By exploring the content and correlation of inorganic elements in soil and Paeoniae Radix Alba in different soil types,as well as the correlation between soil inorganic elements and quality markers of Paeoniae Radix Alba,we could provide reference for the selection of Paeoniae Radix Alba production area and the construction of planting base.The high-quality Paeoniae Radix Alba germplasm types were determined by comparing the interspecific differences and intrspecific stability of existing Paeoniae Radix Alba varieties from Bozhou and Paeoniae Radix Alba varieties introduced to Bozhou from other main producing areas.To provide effective guidance for scientific and reasonable selection of Paeoniae Radix Alba varieties and planting time,we studied the accumulation of quality markers and the changes of main chemical components in their roots of two stable Paeoniae Radix Alba varieties from Bozhou under different planting years.The effects of different fertilization rates on the quality and yield of Paeoniae Radix Alba were designed to explore the effects of different fertilization rates on the quality of Paeoniae Radix Alba,in order to achieve the goal of reducing the application of medicinal fertilizer and increasing the efficiency.Methods:1.Using electrothermal digestion and inductively coupled plasma mass spectrometry(ICP-MS),45Sc,73Ge,115In,209Bi as internal standards,The contents of 18 trace elements including 27Al,48Ti,55Mn,57Fe,59Co,60Ni,63Cu,66Zn,71Ga,75As,82Se,85Rb,88Sr,95Mo,111Cd,133Cs,137Ba and 208Pb in Paeoniae Radix Alba and soil samples were determined.The trace elements in soil and Paeoniae Radix Alba and the relationship between trace elements and Q-marker were analyzed by chemometric method and Person analysis.2.The content of Q-marker in Paeoniae Radix Alba samples of 8 germplasm samples was determined by high performance liquid chromatography(HPLC)on a C18column with 0.1%water phosphoric acid and acetonitrile as the mobile phase and gradient elution.The quality of different germplasm samples was evaluated by combining chemometry and separation distance method.3.The age of the roots of two Paeoniae Radix Alba varieties cultivated in Bozhou was identified by phlorogsanol staining.Combined with the content of Q-marker,the quality of the roots of the same plant with different ages was evaluated,and the influence of cultivation age on the quality of the two Paeoniae Radix Alba varieties was explored.4.An experimental plot was set up in Qiaocheng District of Bozhou City to cultivate two Paeoniae Radix Alba varieties from Bozhou.The application rate of potassium sulfate compound fertilizer(15-15-15)was used as a variable.After three years of cultivation,the yield and quality were measured to evaluate the effect of fertilizer application on the quality and yield of Paeoniae Radix Alba.Results:1.Soil type was significantly positively correlated with the content of Cr,Ga,Rb and Cs in soil,and significantly negatively correlated with the content of Al,but not correlated with the content of Q-marker in Paeoniae Radix Alba.The results showed that the content of Fe was the highest,followed by Zn,Al,Ti and Sr,and the contents of the five heavy metals in the Chinese Pharmacopoeia were all in line with or far below the limited standard.Among the 18 inorganic elements in Paeoniae Radix Alba,Zn had the largest enrichment coefficient,followed by Cu and Sr.Fe,Sr,Zn and Cu in Paeoniae Radix Alba were correlated with Mn,Fe,Co,Ni,Cu,Cd and Ti in soil.The contents of Ni in Paeoniae Radix Alba,Al,As,Sr,Mo and other inorganic elements in soil were positively correlated with Q-marker,while Cu and Zn in medicinal materials and Mn in soil were negatively correlated with Q-marker.2.The fingerprints of Q-Marker components of Paeoniae Radix Alba in different germplasm were established.The similarity evaluation results showed that the similarity of 40 batches of samples was between 0.892 and 1.The Q-Marker content of Paeoniae Radix Alba varies greatly among different germplasm,and the difference mainly comes from the difference of germplasm,that is,the difference of genetic material of the original germplasm.The results of cluster analysis were consistent with those of the fingerprint similarity evaluation,indicating that there were both similarities and differences among the eight germplasm Paeoniae Radix Alba samples.PCA results showed that all the eight components of Q-Marker used in this study could be used as characteristic chemical components affecting the Paeoniae Radix Alba quality of different germplasm.The results of OPLS-DA showed that the content of Paeoniflorin,Albiflorin and PGG varied greatly among batches,which could be used as characterization compounds for Paeoniae Radix Alba differential analysis of different germplasm.TOPSIS results showed that Paeoniae Radix Alba from Yuncheng,Shanxi Province had high quality and stable quality,while the quality of“Zahua Shaoyao”was poor.3.The results showed that there was a strong consistency between Paeoniae Radix Alba of different cultivars and different growth years,and there was a positive correlation between Paeoniae Radix Alba and growth years.The evaluation of cluster analysis classified the 5-year"Pubang"as the same as the 4-year"Xiantiao"and 4-year"Pubang".All the eight components of Q-marker could be used as the characteristic components of Paeoniae Radix Alba in different years,with a cumulative contribution rate of 86.73%.Quantitative analysis of Q-marker showed that the content of paeoniflorin and albiflorin was high in all samples.The contents of gallic acid,paeoniflorin,albiflorin and benzoyl paeoniflorin in"Pubang"were significantly negatively correlated with the growth age,and decreased with the increase of growth age.The content of albiflorin and PGG of"Xiantiao"decreased with the increase of age,while other components were higher in the most recent and oldest years,which may be related to the accumulation and transmission of nutrients and should be further studied.4.For"Xiantiao",the use of 12.375kg/667m~2compound fertilizer could improve the yield to a certain extent,and the application of 8.25kg/667m~2and 4.125kg/667m~2fertilizer had no significant difference with no fertilizer.The yield of"Pubang"under the condition of fertilization was not as good as that without fertilization,and the maximum yield was reached at 8.25kg/667m~2of fertilizer.The similarity of Pubang and Xiantiao under different fertilization conditions was the lowest under the lowest and highest fertilization rates,and the similarity of 4.125kg/667m~2and 8.25kg/667m~2was the highest.Cluster analysis could distinguish the samples with or without fertilization,indicating that fertilization rate had an effect on the growth of Paeoniae Radix Alba.When Q-marker was used as the evaluation index,the effect of fertilizer application on the quality of"Xiantiao"was reverse,and the quality of 8.25kg/667m~2was relatively better under the condition of fertilization,which might be related to the significant negative correlation or no correlation between fertilizer application and paeoniflorin and benzoyl paeoniflorin.On the contrary,high-dose fertilization had a significant effect on"Pubang",and the results of significant positive correlation between fertilization rate and gallic acid,oxpaeoniflorin,catechin,albiflorin,and PGG might be the influencing factor.Conclusion:There was no direct relationship between soil type and the content of inorganic elements in soil or the quality of Paeoniae Radix Alba.The correlation between the inorganic elements in soil and Q-marker was greater than that between the elements and the chemical composition of medicinal materials,which might be related to the element promoting or inhibiting the synthesis of principal components.Paeoniae Radix Alba from Yuncheng of Shanxi Province introduced to Bozhou had high quality and stable quality,while that of“Zahua Shaoyao”was poor.The results suggested that the germplasm from Yuncheng of Shanxi Province could provide materials for the screening and optimization of Paeoniae Radix Alba germplasm resources in Bozhou.The effective ingredients of"Pubang"decreased from 4 to 5 years old,so it was considered that the planting age of"Pubang"should not exceed 5 years.The rhizome quality of"Xiantiao"in the largest year was cumulative,so it could grow to 5 years to achieve high quality and excellent yield.After the balance of yield and quality,the fertilizer application rate of"Xiantiao"and"Pubang"should be between 8.25kg/667m2 and 12.375kg/667m2,but the specific fertilizer application rate needs further study. |
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